title: Targeted gene delivery by new folate-polycationic amphiphilic cyclodextrin-DNA nanocomplexes in vitro and in vivo.
abstract: OBJECTIVE
  Development and evaluation of a new targeted gene delivery system by first preforming self-assembled nanocomplexes from a polycationic amphiphilic cyclodextrin (paCD) and pDNA and then decorating the surface of the nanoparticles with folic acid (FA).
  The cyclodextrin derivative (T2) is a tetradecacationic structure incorporating 14 primary amino groups and 7 thioureido groups at the primary face of a cyclomaltoheptaose (β-CD) core and 14 hexanoyl chains at the secondary face.
  
  
  CONCLUSIONS
  T2 complexed and protected pDNA (luciferase-encoding plasmid DNA, pCMVLuc) and efficiently mediated transfection in vitro and in vivo with no associated toxicity. The combination of folic acid with CDplexes afforded ternary nanocomplexes (Fol-CDplexes) that enhanced significantly the transfection activity of pCMVLuc in human cervix adenocarcinoma HeLa cells, especially when formulated with 1 μg FA/μg DNA. The observed transfection enhancement was associated to specific folate receptor (FR)-mediated internalization of Fol-CDplexes, as corroborated by employing a receptor-deficient cell line (HepG2) and an excess of free folic acid. The in vivo studies, including luciferase reporter gene expression and biodistribution, indicated that 24h after intravenous administration of the T2-pDNA nanocomplexes, transfection takes part mainly in the liver and partially in the lung. Interestingly, the corresponding Fol-CDplexes lead to an increase in the transfection activity in the lung and the liver compared to non-targeted CDplexes. Folate-CDplexes developed in this study have improved transfection efficiency and although various methods have been used for the preparation of ligand-DNA-complexes, covalent binding is usually needed and insoluble aggregates are formed unless the concentration of the components is minimized. However, the complexes developed by first time in this work were prepared by simple mixing. The synthetic nature of this formulation provides the potential of flexibility in terms of composition and the capability of inexpensive and large-scale production of the complexes. These nanovectors may be an adequate alternative to viral vectors for gene therapy in the future.
authors.affiliation: Department of Pharmacy and Pharmaceutical Technology, School of Pharmacy, University of Navarra, Pamplona, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Aranda
authors.forename: Koldo
authors.initials: K
authors.lastname: Urbiola
authors.forename: Alejandro
authors.initials: A
authors.lastname: Méndez Ardoy
authors.forename: José M
authors.initials: JM
authors.lastname: García Fernández
authors.forename: Carmen
authors.initials: C
authors.lastname: Ortiz Mellet
authors.forename: Conchita Tros
authors.initials: CT
authors.lastname: de Ilarduya
mesh_terms: D003505:Cyclodextrins / Q000737:chemistry; D004247:DNA / Q000008:administration & dosage*; D005260:Female; D058973:Folate Receptors, GPI-Anchored / Q000378:metabolism; D005492:Folic Acid / Q000737:chemistry*; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods*; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D008099:Liver / Q000378:metabolism; D008156:Luciferases / Q000235:genetics; D008168:Lung / Q000378:metabolism; D053758:Nanoparticles; D010957:Plasmids; D011073:Polyamines / Q000737:chemistry; D000071228:Polyelectrolytes; D014018:Tissue Distribution; D014162:Transfection
chemical_list: D003505:Cyclodextrins; D004247:DNA; D058973:Folate Receptors, GPI-Anchored; D005492:Folic Acid; D008156:Luciferases; D011073:Polyamines; D000071228:Polyelectrolytes
keywords: Folate receptors; Folic acid; Gene delivery; Nanomedicine; Pharmaceutics; Polycationic amphiphilic cyclodextrins